ID Untitled standard; circular DNA; 6998 BP. XX XX DT 16-SEP-1995 XX XX XX XX XX RN 1 RA Smith,D.B. and Johnson,K.S. RT Single-step purification of polypeptides expressed in Escherichia RT coli as fusions with glutathione S-transferase RL Gene 67, 31-40 (1988) XX RN [2] RA Guan, K-L., Dixon, J.E. RT Eukaryotic proteins expressed in Escherichia coli: an imporived RT thrombin cleavage purification procedure of fusion proteins with RT glutathione S-transferase RL Anal. Biochem 1991;192:262-267 XX CC Ww DO NOT have a good record for how this plasmid was constructed. CC The best guess is that Jin Liu used the m1.9Bam.1S primer and T7 CC to PCR the CDS + 3'UTR from m1.9pBS and ligated the BamH1-Xba1 CC fragment in frame with GST in pGEX.KG XX XX FH Key Location/Qualifiers FH FT CDS 258..1454 FT /note="GST+ mouse AIRAP FT /codon_start=1" FT primer 258..275 FT /note="GST.Xho.3S" FT primer 259..276 FT /note="GST.Xho.1S" FT primer 690..708 FT /note="GEX-KG-2 (Ed Skolnik)" FT primer 869..891 FT /note="GST.Seq.1S ordered 12.03.04 (pGEX.5'seq FT pharmacia)" FT misc_feature 918..935 FT /note="Thrombin cleavage site" FT primer 931..954 FT /note="m1.9Bam.1S" FT 3'UTR 1452..1508 FT /note="mAIRAP 3'UTR" XX SQ Sequence 6998 BP; 1699 A; 1747 C; 1810 G; 1742 T; AGCTTATCGA CTGCACGGTG CACCAATGCT TCTGGCGTCA GGCAGCCATC GGAAGCTGTG GTATGGCTGT GCAGGTCGTA AATCACTGCA TAATTCGTGT CGCTCAAGGC GCACTCCCGT TCTGGATAAT GTTTTTTGCG CCGACATCAT AACGGTTCTG GCAAATATTC TGAAATGAGC TGTTGACAAT TAATCATCGG CTCGTATAAT GTGTGGAATT GTGAGCGGAT AACAATTTCA CACAGGAAAC AGTATTCATG TCCCCTATAC TAGGTTATTG GAAAATTAAG GGCCTTGTGC AACCCACTCG ACTTCTTTTG GAATATCTTG AAGAAAAATA TGAAGAGCAT TTGTATGAGC GCGATGAAGG TGATAAATGG CGAAACAAAA AGTTTGAATT GGGTTTGGAG TTTCCCAATC TTCCTTATTA TATTGATGGT GATGTTAAAT TAACACAGTC TATGGCCATC ATACGTTATA TAGCTGACAA GCACAACATG TTGGGTGGTT GTCCAAAAGA GCGTGCAGAG ATTTCAATGC TTGAAGGAGC GGTTTTGGAT ATTAGATACG GTGTTTCGAG AATTGCATAT AGTAAAGACT TTGAAACTCT CAAAGTTGAT TTTCTTAGCA AGCTACCTGA AATGCTGAAA ATGTTCGAAG ATCGTTTATG TCATAAAACA TATTTAAATG GTGATCATGT AACCCATCCT GACTTCATGT TGTATGACGC TCTTGATGTT GTTTTATACA TGGACCCAAT GTGCCTGGAT GCGTTCCCAA AATTAGTTTG TTTTAAAAAA CGTATTGAAG CTATCCCACA AATTGATAAG TACTTGAAAT CCAGCAAGTA TATAGCATGG CCTTTGCAGG GCTGGCAAGC CACGTTTGGT GGTGGCGACC ATCCTCCAAA ATCGGATCTG GTTCCGCGTG GATCCATAAT GGAGTTTCCT GACTTGGGGA AGCACTGTTC AGAACCAACT TGCAAACAGC TAGATTTTCT GCCAATAACA TGCGACGCCT GTAAACAAGA TTTCTGTAAA GACCATTTTT CCTACGTGGG TCATAAGTGT CCCTTTGCGT TCAAGAAGGA TGTGCAGGTG CCTGTGTGTC CTCTCTGCAA TGCCCCCATC CCTGTGAAGA GAGGTGAGAT CCCAGATGTG GTGGTCGGCG AGCACATGGA CAGAGATTGC ACCTTTCACC CTGGGAGGAA CAGAAACAAG GTTTTCACAC ATCGCTGCTC CAAAGAGGGA TGCAGGAAGA AGGAGATGCT GCAGCTGGCG TGTGCTCAGT GCCATGGCAA CTTCTGCATT CAGCACAGGC ATCCTCTGGA CCACAATTGC CAGGCTGGGA GCAGCTCAGC CAGCAGAGGG AGGACTTCTA CATCCAGAGC TGCTGAGCAG AAGCCATCGG GAGTCAGTTG GCTGGCCCAG CGGCTCAGGC GCACAGTGAA GTGACAGCGA GTCTCCTGTG GGTGCCACAC ACCAGCTCCT TCCTGTTGGA TTTTGATCTT GGGACGTGTC CTCCAACACT TTTCCTGCTT CTTCTGCATC TGAGTCTTTA TAGAAGAGTC TCCCATTGAC TGTTCAAGTT ACGGTGAATT GATGGCTCAG TGGGGAAAGC ACTTGCCCCA CAGGCATGAG GACCTGAGTT CATCCCCAGA ACTGGATGTG ACTGGTGTAC ACTTGTCATC TCAGCACTTG GGAGGTGGTG AGAGGAAAAT CTCCAGGGCT TTCTGACCAG CAGCCTAGCC ATATTGGAAA GTTCCAGGCC AGAAAGACGC TGCCTCAAAA ACCAAGGTGG ACACTTTCTG TGGAGGGACA TCAGAAGTTC ACCTCTGGCC TCACACCCAC ACCCACACAC CCACTTGTGT ATACACACAC ACACACACAC ACACACACAC ACACACACAC ACACACACCC GAGGCTGTCC AGCAACTTAC TTGGGGTGGG TTAGGTTACC ATAGCAAACA GCTGCAGGGT TCAAGGCCTG AGGACTTGCT TAACTCTCCT CAGTGGTGGC TTTGGCCTGT GCTAAACCTC TCCAGGACTT GGTATTCCAG AAAATTCTGT CCTGTGACAG AAAAGAACCA CCGTCTGCTC TGTATTCTCC CCTCCATGGC TTTTGTAATA AGGTGCCTTT GGGCACAGTC TACTCATGGT CCCCAAGAAC AAGAGTGGAC GGGGTGCCCT TTCCTTACCT ACTGGCTGGG GATGCTATGG AGGAAGGGTG TGCACTTGGC CTGCAGTGAT GTCTCCAAGG TTGACTTAAG CAACCAGCCT CACCATGCCA GGACTCCGCA TGCCTGGCTT CAGGCTGTTT TGTGGTTTGA GGCAGTGTCT CACTGTGGAG CCCTGGCTGT CATGGAACTC ACCCTGTAGA CCACACTGGC CTTGAACTCA GTGATCCCCC TGACTCTGCC TCCCAAGCGT GGATTCACTT TTAACTCCCA AGAAGAGCAC CAAGAAATGA GAGCCATTTC AGAAATGTCT GGTTTTGCCA TAGAAGCTTC TCCTCTTCGT GCCGAATCCC CATGGCAGCA GGCTGAGGGT GTCCTCAGCA CTTTGTGGAG CAGAAACCCA GACAGAAGAA AAGCAGTGGT TCTGGGCCTT GGGGTGCAGC CACCATCACT CCTGTGCCAC CTGTGTGGCC TGAACCATGT CTACCCCCAA ATGGCCATGG CAGTGTAGGG GAAGATGCTT GGGGTACTCT GGTGACCTGT ATAAGCGTTT ACCCTCACTT TCCAGGAGCT TGGCTATTAG AGATGACATG GGACCAACCA CCCAGCGGGT TTTGACTTTC GCCCTTAGTT CTGACAGCCT TTGGGACAGT TAACTGCATG AATTCCATAC ATAGCAGCTT TGTGCAGGGT TCGTGGCCTC TAGCTGTCCT GGGAGGCAGG GGTGGGTTCT ACTCACTCTG TTCTCTAAGC CAGTGCCTCT CGTCACATCA GTCCAGCTCT AGACTCCATG GGTCGACTCG AGCTCAAGCT TAATTCACGT GACTGACTGA CGATCTGCCT CGCGCGTTTC GGTGATGACG GTGAAAACCT CTGACACATG CAGCTCCCGG AGACGGTCAC AGCTTGTCTG TAAGCGGATG CCGGGAGCAG ACAAGCCCGT CAGGGCGCGT CAGCGGGTGT TGGCGGGTGT CGGGGCGCAG CCATGACCCA GTCACGTAGC GATAGCGGAG TGTATAATTC TTGAAGACGA AAGGGCCTCG TGATACGCCT ATTTTTATAG GTTAATGTCA TGATAATAAT GGTTTCTTAG ACGTCAGGTG GCACTTTTCG GGGAAATGTG CGCGGAACCC CTATTTGTTT ATTTTTCTAA ATACATTCAA ATATGTATCC GCTCATGAGA CAATAACCCT GATAAATGCT TCAATAATAT TGAAAAAGGA AGAGTATGAG TATTCAACAT TTCCGTGTCG CCCTTATTCC CTTTTTTGCG GCATTTTGCC TTCCTGTTTT TGCTCACCCA GAAACGCTGG TGAAAGTAAA AGATGCTGAA GATCAGTTGG GTGCACGAGT GGGTTACATC GAACTGGATC TCAACAGCGG TAAGATCCTT GAGAGTTTTC GCCCCGAAGA ACGTTTTCCA ATGATGAGCA CTTTTAAAGT TCTGCTATGT GGCGCGGTAT TATCCCGTGT TGACGCCGGG CAAGAGCAAC TCGGTCGCCG CATACACTAT TCTCAGAATG ACTTGGTTGA GTACTCACCA GTCACAGAAA AGCATCTTAC GGATGGCATG ACAGTAAGAG AATTATGCAG TGCTGCCATA ACCATGAGTG ATAACACTGC GGCCAACTTA CTTCTGACAA CGATCGGAGG ACCGAAGGAG CTAACCGCTT TTTTGCACAA CATGGGGGAT CATGTAACTC GCCTTGATCG TTGGGAACCG GAGCTGAATG AAGCCATACC AAACGACGAG CGTGACACCA CGATGCCTGC AGCAATGGCA ACAACGTTGC GCAAACTATT AACTGGCGAA CTACTTACTC TAGCTTCCCG GCAACAATTA ATAGACTGGA TGGAGGCGGA TAAAGTTGCA GGACCACTTC TGCGCTCGGC CCTTCCGGCT GGCTGGTTTA TTGCTGATAA ATCTGGAGCC GGTGAGCGTG GGTCTCGCGG TATCATTGCA GCACTGGGGC CAGATGGTAA GCCCTCCCGT ATCGTAGTTA TCTACACGAC GGGGAGTCAG GCAACTATGG ATGAACGAAA TAGACAGATC GCTGAGATAG GTGCCTCACT GATTAAGCAT TGGTAACTGT CAGACCAAGT TTACTCATAT ATACTTTAGA TTGATTTAAA ACTTCATTTT TAATTTAAAA GGATCTAGGT GAAGATCCTT TTTGATAATC TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC TTGCAAACAA AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTCCTTCTA GTGTAGCCGT AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT CTGCTAATCC TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG GACTCAAGAC GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC ACACAGCCCA GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA TGAGAAAGCG CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG GTCGGAACAG GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT CCTGTCGGGT TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG CGGAGCCTAT GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC ACATGTTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC GCCTTTGAGT GAGCTGATAC CGCTCGCCGC AGCCGAACGA CCGAGCGCAG CGAGTCAGTG AGCGAGGAAG CGGAAGAGCG CCTGATGCGG TATTTTCTCC TTACGCATCT GTGCGGTATT TCACACCGCA TAAATTCCGA CACCATCGAA TGGTGCAAAA CCTTTCGCGG TATGGCATGA TAGCGCCCGG AAGAGAGTCA ATTCAGGGTG GTGAATGTGA AACCAGTAAC GTTATACGAT GTCGCAGAGT ATGCCGGTGT CTCTTATCAG ACCGTTTCCC GCGTGGTGAA CCAGGCCAGC CACGTTTCTG CGAAAACGCG GGAAAAAGTG GAAGCGGCGA TGGCGGAGCT GAATTACATT CCCAACCGCG TGGCACAACA ACTGGCGGGC AAACAGTCGT TGCTGATTGG CGTTGCCACC TCCAGTCTGG CCCTGCACGC GCCGTCGCAA ATTGTCGCGG CGATTAAATC TCGCGCCGAT CAACTGGGTG CCAGCGTGGT GGTGTCGATG GTAGAACGAA GCGGCGTCGA AGCCTGTAAA GCGGCGGTGC ACAATCTTCT CGCGCAACGC GTCAGTGGGC TGATCATTAA CTATCCGCTG GATGACCAGG ATGCCATTGC TGTGGAAGCT GCCTGCACTA ATGTTCCGGC GTTATTTCTT GATGTCTCTG ACCAGACACC CATCAACAGT ATTATTTTCT CCCATGAAGA CGGTACGCGA CTGGGCGTGG AGCATCTGGT CGCATTGGGT CACCAGCAAA TCGCGCTGTT AGCGGGCCCA TTAAGTTCTG TCTCGGCGCG TCTGCGTCTG GCTGGCTGGC ATAAATATCT CACTCGCAAT CAAATTCAGC CGATAGCGGA ACGGGAAGGC GACTGGAGTG CCATGTCCGG TTTTCAACAA ACCATGCAAA TGCTGAATGA GGGCATCGTT CCCACTGCGA TGCTGGTTGC CAACGATCAG ATGGCGCTGG GCGCAATGCG CGCCATTACC GAGTCCGGGC TGCGCGTTGG TGCGGATATC TCGGTAGTGG GATACGACGA TACCGAAGAC AGCTCATGTT ATATCCCGCC GTCAACCACC ATCAAACAGG ATTTTCGCCT GCTGGGGCAA ACCAGCGTGG ACCGCTTGCT GCAACTCTCT CAGGGCCAGG CGGTGAAGGG CAATCAGCTG TTGCCCGTCT CACTGGTGAA AAGAAAAACC ACCCTGGCGC CCAATACGCA AACCGCCTCT CCCCGCGCGT TGGCCGATTC ATTAATGCAG CTGGCACGAC AGGTTTCCCG ACTGGAAAGC GGGCAGTGAG CGCAACGCAA TTAATGTGAG TTAGCTCACT CATTAGGCAC CCCAGGCTTT ACACTTTATG CTTCCGGCTC GTATGTTGTG TGGAATTGTG AGCGGATAAC AATTTCACAC AGGAAACAGC TATGACCATG ATTACGGATT CACTGGCCGT CGTTTTACAA CGTCGTGACT GGGAAAACCC TGGCGTTACC CAACTTAATC GCCTTGCAGC ACATCCCCCT TTCGCCAGCT GGCGTAATAG CGAAGAGGCC CGCACCGATC GCCCTTCCCA ACAGTTGCGC AGCCTGAATG GCGAATGGCG CTTTGCCTGG TTTCCGGCAC CAGAAGCGGT GCCGGAAAGC TGGCTGGAGT GCGATCTTCC TGAGGCCGAT ACTGTCGTCG TCCCCTCAAA CTGGCAGATG CACGGTTACG ATGCGCCCAT CTACACCAAC GTAACCTATC CCATTACGGT CAATCCGCCG TTTGTTCCCA CGGAGAATCC GACGGGTTGT TACTCGCTCA CATTTAATGT TGATGAAAGC TGGCTACAGG AAGGCCAGAC GCGAATTATT TTTGATGGCG TTGGAATT //